CN103565928A - Guava hypoglycemic active component, and preparation method and use thereof - Google Patents
Guava hypoglycemic active component, and preparation method and use thereof Download PDFInfo
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Abstract
The invention provides a guava hypoglycemic active component, and a preparation method and use thereof. The method comprises the following steps: crushing a raw material of guava leaves; carrying out reflux extraction by 8-10fold by volume of 50% ethanol; carrying out vacuum concentration below 60 DEG C; dispersing the obtained ethanol extract into water; fully extracting by adopting ethyl acetate after petroleum ether is extracted; carrying out vacuum concentration below 60 DEG C; filtering the obtained ethyl acetate extract by a 60-200mesh polyamide chromatographic column; eluting ethanol water from low to high in a gradient manner; collecting a component which is eluted by ethanol of which the concentration is 30-50%, continuing to filter by the 60-200mesh polyamide chromatographic column after vacuum concentration below 60 DEG C; carrying out isocratic elution by 30% ethanol after eluting by 10% ethanol; carrying out vacuum concentration on eluent below 60 DEG C, and recrystallizing by a 70% ethanol solvent, so as to obtain the hypoglycemic active component. The method disclosed by the invention is simple and convenient; chemical components of an effective part are clear in composition, and good in chemical stability. By adopting the guava hypoglycemic active component disclosed by the invention, alloxan-induced hyperglycemia can be significantly reduced, and the guava hypoglycemic active component can be applied to preparation of hypoglycemic medicaments or health-care products.
Description
Technical field
The present invention relates to a kind of health-caring product capable of reducing blood sugar field, particularly a kind of Fructus psidii guajavae immaturus hypoglycemic activity component, Preparation method and use.
Background technology
Folium Granati is used for preventing as medication among the people and the existing long history for the treatment of diabetes, there are a large amount of bibliographical informations of process of preventing and treating type 2 diabetes mellitus patient disease about Folium Psidii Guajavae, and a large amount of clinical trials proves its determined curative effect, therefore domestic and international many scholars have launched a large amount of research to its hypoglycemic effect, but up to the present, in Folium Psidii Guajavae, blood-sugar decreasing active and mechanism of action are not yet definitely confirmed.Chinese patent application disclosed " a kind of pharmaceutical composition or health product with effect of lowering blood sugar " (201210505096.8), a kind of hypoglycemic drug being comprised of Fructus Momordicae charantiae extract, Fructus Canrii Albi extract, Cortex Cinnamomi extract, guava extract, Radix Et Rhizoma Fagopyri Tatarici extract, astaxanthin, phylloxanthin, yeast chromium, xylitol is provided, but extracting method and the concrete composition of itself and undeclared guava extract wherein.Folium Psidii Guajavae resource is very abundant in China, if Folium Psidii Guajavae is goed deep into systematic research, determine its blood sugar reducing function material base, identify the chemical constitution of its blood sugar lowering monomer, bright its mechanism of action of analysing, and develop on this basis and prevent and treat hypoglycemic medicine or the functional food that development occurs type 2 diabetes mellitus, not only can make full use of Folium Psidii Guajavae resource, but also can reduce diabetes patient misery, create larger economic benefit and social benefit.
Summary of the invention
The object of the present invention is to provide a kind of hypoglycemic activity component of extracting from Folium Psidii Guajavae, can be effective to prepare hypoglycemic medicine or functional food, for type 2 diabetes mellitus patient's control, for hyperglycemic patients provides healthy.The present invention also provides a kind of preparation method of Fructus psidii guajavae immaturus hypoglycemic activity component and this Fructus psidii guajavae immaturus hypoglycemic activity component in the purposes of preparing aspect hypoglycemic medicine and health product.
For achieving the above object, embodiment of the present invention are: a kind of Folium Psidii Guajavae hypoglycemic activity component, is prepared from by the method comprising the following steps:
(1) by Folium Psidii Guajavae raw material, pulverizing, 8-10 times of volume 50% alcohol reflux 2 times, 60 ℃ of following concentrating under reduced pressure of extracting solution obtain ethanol extraction;
(2) ethanol extraction is scattered in water, adopts ethyl acetate fully to extract after petroleum ether extraction, combined ethyl acetate extract, and 60 ℃ of following concentrating under reduced pressure obtain acetic acid ethyl ester extract;
(3) acetic acid ethyl ester extract is crossed 60-200 order polyamide chromatography post, the ethanol water that is 0%-95% by concentration is gradient elution from low to high, collect the component with 30%-50% concentration ethanol eluting, after 60 ℃ of following concentrating under reduced pressure, continued 60-200 order polyamide chromatography, after 10% ethanol elution, 30% ethanol isocratic elution, collects eluent, and 60 ℃ of following concentrating under reduced pressure, 70% alcohol solvent recrystallization obtain hypoglycemic activity component.
In step (3), during polyamide chromatography column chromatography, extract and adsorbent weight are than being 1:10～1:30.During ethanol gradient elution, concentration of alcohol is followed successively by: 0%, 10%, 30%, 50%, 70%, 95%.
Measure by analysis and determine that this blood sugar lowering component contains five kinds of flavone monomeric compounds, being respectively hyperin, isoquercitrin, Quercetin-3-O-β-D-xylopyranoside, guaijaverin, auicularin.Five kinds of content of monomer sums 50%～95%, guaijaverin and auicularin content sum 25%～75%.
The inventive method is easy, and live part chemical composition forms clear, and stable chemical nature is good, has opened up the new situation of Folium Psidii Guajavae function of polysaccharide application, is an innovation in functional food field.Through whole blood sugar lowering zoopery, show, product of the present invention can significantly reduce the hyperglycemia of alloxan induction, can be used for preparing hypoglycemic drug or health product.
Accompanying drawing explanation
Fig. 1 is glucose content comparison in Folium Psidii Guajavae runic thing adipocyte culture liquid (noting: * p<0.01, #p<0.05vs matched group);
Fig. 2 is glucose content comparison in Folium Psidii Guajavae runic thing adipocyte culture liquid (noting: * p<0.01, #p<0.05vs matched group);
Fig. 3 is glucose content comparison in column chromatography 1 each component adipocyte culture liquid (noting: * p<0.01, #p<0.05vs matched group);
Fig. 4 is C1-D chromatography result.
The specific embodiment
Below in conjunction with practical situation, the specific embodiment of the present invention is elaborated.
Embodiment 1:
Take JIUYUE Folium Psidii Guajavae raw material 1.2kg, 10L50% alcohol reflux 2 times, each 1 hour, 60 ℃ of following concentrating under reduced pressure of extracting solution obtained ethanol extraction, ethanol extraction is scattered in water, when petroleum ether extraction shoals to solution colour, change ethyl acetate extraction 6 times into, to last extract very slight color, merge concentrated extract and obtain acetic acid ethyl ester extract 35.6g, extract is crossed 100 order polyamide column 400g, washing, 4 times of column volumes of each eluting of 10% ethanol, 50% 8 times of ethanol elutions column volume, concentrate to obtain 10.3g solid, after 200 order polyamide columns, 10% 3 times of ethanol elutions column volume, 10 times of column volumes of rear 30% ethanol isocratic elution, concentrated, 70% alcohol crystal obtains hypoglycemic activity component 4.97g, liquid phase measure and calculation contains auicularin 1.19g, guaijaverin 1.03g, Quercetin-3-O-β-D-xylopyranoside 0.57g, isoquercitrin 0.19g, hyperin 0.45g.
Embodiment 2:
Take Folium Psidii Guajavae raw material 20kg in October, 200L50% alcohol reflux 2 times, each 2 hours, 60 ℃ of following concentrating under reduced pressure of extracting solution obtained ethanol extraction, ethanol extraction is scattered in water, when petroleum ether extraction shoals to solution colour, change ethyl acetate extraction 10 times into, to last extract very slight color, merge concentrated extract and obtain acetic acid ethyl ester extract 519.2g, extract is crossed 80 order polyamide columns, washing, the abundant eluting of 10% ethanol, 50% 10 times of ethanol elutions column volume, concentrate to obtain 177.8g solid, after 100 order polyamide columns, 10% 6 times of ethanol elutions column volume, rear 30% ethanol isocratic elution, concentrated, 70% alcohol crystal obtains hypoglycemic activity component 103.54g, liquid phase measure and calculation contains auicularin 19.72g, guaijaverin 23.49g, Quercetin-3-O-β-D-xylopyranoside 4.56g, isoquercitrin 1.38g, hyperin 6.95g.
For illustrating, verify that the present invention reduces the effect of hyperglycemia, spy has done following blood sugar lowering experiment.
1. the foundation of glucose consumption blood sugar lowering fat cell model
The disconnected neck of male SD rat is put to death, and the aseptic abdomen of opening is got the outer white adipose tissue of epididymis, rejects subsidiary blood vessel, shreds to about 1mm
3size, adds type i collagen enzymic digestion 1h, collects upper strata adipose cell, adds appropriate Hanks buffer washed cell to obtain Primary adipocyte three times.In 96 orifice plates, set up 200 μ L reaction systems: concentration is 2 * 10
6cells/mL cell suspension 100 μ L hyclone+40, μ L+30 μ L6mg/mL glucose+30 μ L sample solutions (comprising each extract or each column chromatography elution fraction), with saturated humidity, 37 ℃, 5%CO
2cultivate reaction 30min, every hole adds 1 μ L2mmol/L epinephrine solution again, then cultivates 3h, ice bath cessation reaction with similarity condition.Adopt micro-glucose oxidase method colour developing, reading in microplate reader, measures and calculates each hole concentration of glucose, and relatively calculates with matched group the consumption concentration of respectively organizing glucose, the ingestion of glucose ability of evaluating thus each extract or each column chromatography component, specific formula for calculation is as follows:
glucose-uptake?level=C
ctrl-C
compound(mmol/L)
Wherein, C
ctrlfor the concentration of matched group glucose, C
compoundthe concentration of glucose in residual culture medium after 3 hours cultivate for sample or the separated effective ingredient obtaining.In this model, with 0.1%DMSO Hanks buffer, do blank group, insulin is done positive control.
2. the active results and analysis of Folium Psidii Guajavae crude extract
Set up cell model, detect respectively crude extract WE (water extract), 50AE (50% ethanol extraction), 85AE (85% ethanol extraction), EAE (ethyl acetate extract), five components of PE (ligroin extraction) are under 1.6mg/mL, 1.2mg/mL and 0.8mg/mL in concentration, impact on adipose cell glucose metabolism, the results are shown in Figure 1.
Fig. 1 demonstration, different component has a certain impact to glucose content in adipocyte culture liquid.Along with the increase of insulin concentration, in culture fluid, glucose content reduces gradually, shows dose-dependence.In 5 sample component, contrast matched group, 50AE can significantly reduce glucose content in Cell sap, and in culture fluid, glucose content is minimum, and is dose-dependence, and when 50AE concentration is during at 1.6mg/mL, the insulin of hypoglycemic effect and 8U/mL is suitable.Whole hypoglycemic effect relatively sorts: 50AE>WE>85AE>E AE>PE, from sequence, observe, when polarity is 50% ethanol, its extract has the strongest hypoglycemic activity, when the polarity of extracting solution is to increasing or dwindling while changing, the hypoglycemic activity of extract reduces along with change in polarity, and activity shows as polarity dependence.
3. the active results and analysis of Folium Psidii Guajavae extract
50AE sample employing polarity from low to high solvent system solvent extraction obtains respectively petroleum ether extract (PS), acetic acid ethyl ester extract (EAS).After ethyl acetate extraction, emit remaining aqueous solution and emulsion layer, concentrating under reduced pressure, respectively called after WS and EL.With Petroleum ether extraction EAS extractum, filter, by a small amount of method of repeatedly extracting, until filtrate clarification is colourless, filtrate is merged, concentrating under reduced pressure obtains P-EAS, and remaining filtering residue is R-EAS.Utilize cell model, detect respectively extract EL, WS, R-EAS, tetra-components of P-EAS are under 1.2mg/mL and 0.8mg/mL and 0.6mg/mL in concentration, the impact on adipose cell glucose metabolism, and add the contrast of 50AE component.The results are shown in Figure 2.
Fig. 2 shows, the impact of each extracted component on glucose content in adipocyte culture liquid.Because 50AE component is crude extract, composition is more and complicated than extract, and its activity significantly shows as dose-dependence, so during design 50AE contrast, grade of its concentration ratio extraction object height.In 4 extracted components, contrast matched group, R-EAS can significantly reduce glucose content in Cell sap, in culture fluid, glucose content is minimum, and be dose-dependence, and when R-EAS concentration is during at 1.2mg/mL, hypoglycemic effect is stronger than the 50AE of the insulin of 8U/mL and 1.6mg/mL.Whole hypoglycemic effect relatively sorts: R-EAS>EL>WS>P-EAS.Analyze each component extracting liquid, extracted component extract polarity sequence used is: R-EAS>EL>WS>P-EAS (EL is emulsion layer, so polarity is between water and ethyl acetate).From hypoglycemic effect and polarity sequence, observe, the hypoglycemic activity of extract is relevant to its change in polarity, shows as equally polarity dependence.
4. column chromatography 1 each composition activity results and analysis
R-EAS carries out polyamide column chromatography, obtains 11 components, respectively called after C1-A-C1-K.Utilize under the condition that cell model is 0.6mg/mL to 11 components in concentration, carry out hypoglycemic activity detection.And the R-EAS component that adds 1.2mg/mL compares.The results are shown in Figure 3.
Fig. 3 shows the impact of each column chromatography component on glucose content in adipocyte culture liquid.R-EAS component is extract, the complicated components of composition after than column chromatography, and inert matter abundant species and content are high, in addition, because its activity shows as dose-dependence, so during design R-EAS contrast, its concentration ratio extract doubles.In 11 column chromatography components, contrast matched group, C1-C, C1-D, C1-E show very strong hypoglycemic activity, can significantly reduce glucose content in Cell sap, and in culture fluid, glucose content is minimum with three.From culture fluid, glucose content whole observation is analyzed, and 11 components are parabolic shape to its impact, and wherein take C1-C, C1-D, C1-E is parabolical the lowest point, and both sides increase progressively.In column chromatography process, eluent is to change and carry out gradient elution according to solution polarity.Therefore can tentatively judge, after column chromatography purification, the hypoglycemic activity of active component shows as polarity dependence equally.
In addition in the strongest blood sugar lowering component C1-C, C1-D, C1-E, yield with C1-D is high, and C1-C, C1-E yield are less, liquid chromatographic detection analysis, three contained chemical compositions of the strongest blood sugar lowering component are almost completely consistent, therefore be merged into a component, carry out later separation analysis.
5. the reversed-phase liquid chromatography analysis of " C1-D "
" C1-D " is dissolved in methanol, crosses 0.22 μ m microporous filter membrane, get 5 μ L and inject liquid chromatograph, 17% acetonitrile 1mL/min Gradient elution, 257nm detects analysis, and the following Fig. 4 of chromatograph shows.
By Fig. 4, shown, in blood sugar lowering best composition " C1-D ", contain altogether 5 kinds of monomeric compounds, and having adopted preparative liquid chromatography analysis to get whole 5 kinds of monomeric compounds, 1-peak, peak 5 is respectively hyperin, isoquercitrin, Quercetin-3-O-β-D-xylopyranoside, guaijaverin, auicularin.Meanwhile, area normalization method shows, 5 kinds of content of monomer total contents reach 90%, and guaijaverin content is 28%, auicularin content 33%, and guaijaverin and auicularin content sum reach 63%.
6. prepare liquid phase separation purification and Structural Identification
Adopt the monomer chemical composition containing in preparation liquid phase separation purification C1-D.Chromatographic condition: mobile phase is 32% methanol-water system, preparative hplc post is Shimadzu Shim-pack ODS type C18 post (250mm * 20mm, 5 μ m), detects wavelength 257, and column temperature is room temperature, and flow velocity is 12mL/min.The monomer that separation is obtained adopts magnetic nuclear resonance method to carry out Structural Identification.
6.1 peak 1-hyperins
Peak 1 is yellow powder, and the reaction of hydrochloric acid-magnesium powder is aobvious red, shows that it may be a flavone compound.Analyze its spectral data of mensuration as follows:
1H?NMR(CD
3OD，6in?ppm，J?in?Hz)：6.21(1H，d，J=1.8，H-6)，6.40(1H，d，J=1.8，H-8)，7.83(1H，d，J=2.4，H-2′)，6.87(1H，d，J=8.4，H-5′)，7.58(1H，dd，J=2.4，8.4，H-6′)，5.15(1H，d，J=7.8，H-1″)，3.85(1H，d，J=2.4，H-4").
13C?NMR(CD
3OD，6in?ppm)：158.9(C-2)，135.9(C-3)，179.6(C-4)，105.7(C-4a)，163.0(C-5)，99.9(C-6)，166.0(C-7)，94.8(C-8)，158.5(C-8a)，122.9(C-1′)，117.9(C-2′)，145.9(C-3′)，149.9(C-4′)，116.1(C-5′)，123.0(C-6′)，105.5(C-1″).73.2(C-2")，75.1(C-3″)，70.0(C-4″)，77.2(C-5″)，62.0(C-6").
Above spectral data proof peak 1 is Quercetin 3-O-β-D-galactoside, i.e. hyperin, and structural formula is as follows.
6.2 peak 2-isoquercitrins
1H?NMR(CD
3OD，6in?ppm，J?in?Hz)：6.20(1H，s，H-6)，6.39(1H，s，H-8)，7.71(1H，s，H-2′)，6.88(1H，d，J=7.8，H-5′)，7.58(1H，d，J=7.8，H-6′)，5.23(1H，d，J=7.2，H-1″).
13C?NMR(CD
3OD，6in?ppm)：159.1(C-2)，135.7(C-3)，179.5(C-4)，105.7(C-4a)，163.0(C-5)，99.9(C-6)，166.0(C-7)，94.8(C-8)，158.5(C-8a)，122.1(C-1′)，117.6(C-2′)，145.9(C-3′)，149.9(C-4′)，116.1(C-5′)，123.2(C-6′)，104.4(C-1″)，75.8(C-2")，78.4(C-3″)，71.3(C-4")，78.3(C-5″)，62.6(C-6").
Above spectral data proof peak 2 is Quercetin 3-O-β-D-Glucose glycosides, i.e. isoquercitrin, and structural formula is as follows.
6.3 peak 3-Quercetins-3-O-β-D-xylopyranoside
Peak 3 is yellow powder, and the reaction of hydrochloric acid-magnesium powder is aobvious red, shows that it may be a flavone compound.Analyze its spectral data of mensuration as follows:
1H?NMR(CD
3OD，6in?ppm，J?in?Hz)：6.23(1H，d，J=1.8，H-6)，6.42(1H，d，J=1.8，H-8)，7.61(1H，d，J=1.8，H-2′)，6.88(1H，d，J＝8.4，H-5′)，7.58(1H，dd，J=1.8，8.4，H-6′)，5.16(1H，d，J=7.2，H-1″).
13C?NMR(CD
3OD，6in?ppm)：159.1(C-2)，135.4(C-3)，179.4(C-4)，105.7(C-4a)，162.9(C-5)，100.0(C-6)，166.0(C-7)，94.9(C-8)，158.5(C-8a)，123.0(C-1′)，117.3(C-2′)，146.0(C-3′)，149.8(C-4′)，116.2(C-5′)，123.4(C-6′)，104.5(C-1″)，75.2(C-2")，77.4(C-3″)，71.0(C-4″)，67.1(C-5″).
Above spectral data proof peak 3 Quercetins-3-O-β-D-xylopyranoside, structural formula is as follows.
6.4 peak 4-guaijaverins
1H?NMR(DMSO-d6，6in?ppm，J?in?Hz)：12.6(5-OH)，6.20(1H，d，J=1.8，H-6)，6.40(1H，d，J=1.8，H-8)，7.50(1H，d，J=2.4，H-2′)，6.83(1H，d，J=8.4，H-5′)，7.65(1H，dd，J=2.4，8.4，H-6′)，5.27(1H，d，J=4.8，H-1″).
13C?NMR(DMSO-d6，6in?ppm)：156.3(C-2)，133.8(C-3)，177.5(C-4)，103.9(C-4a)，161.2(C-5)，98.7(C-6)，164.2(C-7)，93.5(C-8)，156.3(C-8a)，120.9(C-1′)，115.8(C-2′)，144.9(C-3′)，148.6(C-4′)，115.4(C-5′)，122.0(C-6′)，101.4(C-1″)，70.7(C-2")，71.7(C-3″)，66.0(C-4″)，64.3(C-5").
Above spectral data proof peak 4 is Quercetin 3-O-α-L arabopyranose glycosides, i.e. guaijaverin, and structural formula is as follows.
6.5 peak 5-auicularins
Peak 5 is yellow crystal, and the reaction of hydrochloric acid-magnesium powder is aobvious red, shows that it may be a flavone compound.Analyze its spectral data of mensuration as follows:
1H?NMR(DMSO-d6，6in?ppm，J?in?Hz)：12.6(5-OH)，6.20(1H，d，J=1.8，H-6)，6.41(1H，d，J=1.8，H-8)，7.47(1H，d，J=2.4，H-2′)，6.84(1H，d，J=9.0，H-5′)，7.55(1H，dd，J=2.4，9.0，H-6′)，5.58(1H，d，J=1.2，H-1″).
13C?NMR(DMSO-d6，6in?ppm)：156.9(C-2)，133.3(C-3)，177.6(C-4)，103.9(C-4a)，161.1(C-5)，98.6(C-6)，164.1(C-7)，93.5(C-8)，156.2(C-8a)，120.9(C-1′)，115.5(C-2′)，145.0(C-3′)，148.4(C-4′)，115.5(C-5′)，121.6(C-6′)，107.8(C-1″)，82.0(C-2")，76.9(C-3″)，85.8(C-4″)，60.6(C-5″).
Above spectral data proof peak 5 is Quercetin 3-O-α-L arabinofuranosyl glycosides, i.e. auicularin, and structural formula is as follows.
7. monomer hypoglycemic activity
Under gnotobasis, 5 kinds of monomers are dissolved in to the Hanks buffer (DMSO final concentration is 0.1%) containing DMSO, cross 0.22 μ m microporous filter membrane, being configured to final concentration is each monomer sample solution of 40 μ mol/L, according to cell model, carry out the analysis of monomer hypoglycemic activity, result is as shown in table 1 below
The impact of 5 kinds of monomers of table 1. Folium Psidii Guajavae on adipose cell glucose uptake amount
The reference value of matched group glucose uptake amount of take is 0, table 1 is all numerical value of adipose cell ingestion of glucose when 40 μ mol/L of 5 kinds of monomers, result shows that compound is the strongest with main effective ingredient guaijaverin in Folium Psidii Guajavae, the short sugared absorbability of auicularin, be respectively 1.23 ± 0.059mmol/L, 1.86 ± 0.194mmol/L, and hyperin, isoquercitrin, Quercetin-3-O-β-D-xyloside blood sugar lowering ability a little less than, this is embodied in these 5 kinds of monomeric compounds, compound polarity is less, and its blood sugar lowering ability is stronger.
8. hypoglycemic activity component impact to mice hyperglycemia level on alloxan
Get 60 of male mices, 25 ± 2 grams of left and right of body weight, get 10 normal controls, after all the other 50 fasting 24h, the modeling of tail vein injection 50mg/kg alloxan suspension, after 5 days, fasting 5h mensuration blood glucose value 10-25mmol/L represents modeling success, be divided at random five groups, the difference large, medium and small dosage of gavage Folium Psidii Guajavae hypoglycemic activity of the present invention component (400mg/kg, 200mg/kg, 100mg/kg) normal saline suspension, metformin suspension 170mg/kg, model group and Normal group gavage isopyknic normal saline solution.Successive administration 4 weeks, after fasting 5h, mouse orbit vein is dehematized, separation of serum, determination of glucose oxidase blood sugar concentration, concrete outcome is as following table 2
Table 2. Fructus psidii guajavae immaturus hypoglycemic activity of the present invention component causes the impact of mice hyperglycemia level on alloxan
# and model group be P<0.05 relatively, and * and model group be P<0.01 relatively
From upper table 2, with Normal group comparison, alloxan modeling group blood glucose significantly raises (P<0.01), and the positive hypoglycemic medicine metformin blood glucose value that can significantly decline, with model group P<0.01 relatively.The present invention obtains Fructus psidii guajavae immaturus hypoglycemic activity component and alloxan is caused to mice hyperglycemia has remarkable downward effect, with model group significance relatively, and embodies dose-effect relationship, and institute's dosage is larger, and hypoglycemic effect is more obvious.
9. sum up
By setting up glucose consumption adipose cell model, under instructing, activity carries out separation and purification, determined the active component in Folium Psidii Guajavae with optimum hypoglycemic effect, by liquid-phase chromatographic analysis and preparative liquid chromatography purification and Spectrum Analysis, determined that in hypoglycemic activity component, 5 kinds of monomers are respectively hyperin, isoquercitrin, Quercetin-3-O-β-D-xylopyranoside, guaijaverin, auicularin; And these 5 kinds of monomer hypoglycemic activities are the strongest with the lower auicularin of polarity, guaijaverin activity.And this hypoglycemic activity thing is carried out to whole blood sugar lowering zoopery, result shows that product of the present invention can significantly reduce the hyperglycemia of alloxan induction.
Claims (6)
1. a Folium Psidii Guajavae hypoglycemic activity component, is characterized in that, by the method comprising the following steps, is prepared from:
(1) by Folium Psidii Guajavae raw material, pulverizing, 8-10 times volume 50% alcohol reflux 2 times, 60 ℃ of following concentrating under reduced pressure of extracting solution obtain ethanol extraction;
(2) ethanol extraction is scattered in water, adopts ethyl acetate fully to extract after petroleum ether extraction, combined ethyl acetate extract, and 60 ℃ of following concentrating under reduced pressure obtain acetic acid ethyl ester extract;
(3) acetic acid ethyl ester extract is crossed 60-200 order polyamide chromatography post, the ethanol water that is 0%-95% by concentration is gradient elution from low to high, collect the component with 30%-50% concentration ethanol eluting, after 60 ℃ of following concentrating under reduced pressure, continued 60-200 order polyamide chromatography, after 10% ethanol elution, 30% ethanol isocratic elution, collects eluent, and 60 ℃ of following concentrating under reduced pressure, 70% alcohol solvent recrystallization obtain hypoglycemic activity component.
2. Folium Psidii Guajavae hypoglycemic activity component according to claim 1, it is characterized in that, contain hyperin, isoquercitrin, Quercetin-3-O-β-D-xylopyranoside, guaijaverin, auicularin, above-mentioned five kinds of content of monomer sums 50%～95%, guaijaverin and auicularin content sum 25%～75%.
3. a preparation method for Folium Psidii Guajavae hypoglycemic activity component as claimed in claim 1, is characterized in that,
By the method comprising the following steps, be prepared from:
(1) by Folium Psidii Guajavae raw material, pulverizing, 8-10 times volume 50% alcohol reflux 2 times, 60 ℃ of following concentrating under reduced pressure of extracting solution obtain ethanol extraction;
(2) ethanol extraction is scattered in water, adopts ethyl acetate fully to extract after petroleum ether extraction, combined ethyl acetate extract, and 60 ℃ of following concentrating under reduced pressure obtain acetic acid ethyl ester extract;
(3) acetic acid ethyl ester extract is crossed 60-200 order polyamide chromatography post, the ethanol water that is 0%-95% by concentration is gradient elution from low to high, collect the component with 30%-50% concentration ethanol eluting, after 60 ℃ of following concentrating under reduced pressure, continued 60-200 order polyamide chromatography, after 10% ethanol elution, 30% ethanol isocratic elution, collects eluent, and 60 ℃ of following concentrating under reduced pressure, 70% alcohol solvent recrystallization obtain hypoglycemic activity component.
4. the preparation method of Folium Psidii Guajavae hypoglycemic activity component according to claim 2, is characterized in that, in step (3), during polyamide chromatography column chromatography, extract and adsorbent weight are than being 1:10-1:30.During ethanol gradient elution, concentration of alcohol is followed successively by: 0%, 10%, 30%, 50%, 70%, 95%.
5. Folium Psidii Guajavae hypoglycemic activity component as claimed in claim 1 is in the purposes of preparing aspect blood sugar reducing health product.
6. Folium Psidii Guajavae hypoglycemic activity component as claimed in claim 1 is in the purposes of preparing blood sugar lowering pharmaceutical preparations.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106117286A (en) * | 2016-06-17 | 2016-11-16 | 成都瑞科信科技有限公司 | A kind of separation stores glycosides and the method for guaijaverin in Folium Psidii Guajavae |
| CN107365336A (en) * | 2017-07-26 | 2017-11-21 | 华南农业大学 | A kind of noval chemical compound extracted from Guava Leaf, preparation method and its usage |
| CN105662946B (en) * | 2016-01-19 | 2019-03-22 | 上海清轩生物科技有限公司 | The preparation of pomegranate rind extract and its application in cosmetics |
| CN111973612A (en) * | 2020-09-15 | 2020-11-24 | 河南大学 | Application of quercetin-3-O-beta-D-galactoside in preparation of medicament for promoting glucose absorption |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007053865A1 (en) * | 2005-11-09 | 2007-05-18 | Omnica Gmbh | Pharmaceutical use of a compound |
| CN101883575A (en) * | 2007-04-27 | 2010-11-10 | 欧博康有限公司 | Guava extract |
| CN102920785A (en) * | 2012-11-27 | 2013-02-13 | 哈药集团中药二厂 | Method for extracting total flavone from guava leaves |
-
2013
- 2013-11-01 CN CN201310528407.7A patent/CN103565928A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007053865A1 (en) * | 2005-11-09 | 2007-05-18 | Omnica Gmbh | Pharmaceutical use of a compound |
| CN101883575A (en) * | 2007-04-27 | 2010-11-10 | 欧博康有限公司 | Guava extract |
| CN102920785A (en) * | 2012-11-27 | 2013-02-13 | 哈药集团中药二厂 | Method for extracting total flavone from guava leaves |
Non-Patent Citations (3)
| Title |
|---|
| AM METWALLY ET AL: "phytochemical investigation and antimicrobial activity of psidium guajava L. leaves", 《PHARMACOGNOSY MAGAZINE》 * |
| 国家中医药管理局专业技术资格考试专家委员会: "《全国中医药专业技术资格考试大纲与细则.中药专业：中级》", 31 December 2011, 中国中医药出版社 * |
| 蔡丹昭: "番石榴叶总黄酮提取及其降血糖机制的研究", 《中国博士学位论文全文数据库 医药卫生科技辑》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105662946B (en) * | 2016-01-19 | 2019-03-22 | 上海清轩生物科技有限公司 | The preparation of pomegranate rind extract and its application in cosmetics |
| CN106117286A (en) * | 2016-06-17 | 2016-11-16 | 成都瑞科信科技有限公司 | A kind of separation stores glycosides and the method for guaijaverin in Folium Psidii Guajavae |
| CN106117286B (en) * | 2016-06-17 | 2018-06-15 | 成都瑞科信科技有限公司 | A kind of method for detaching Guava Leaf Zhong avicularins and guaijaverin |
| CN107365336A (en) * | 2017-07-26 | 2017-11-21 | 华南农业大学 | A kind of noval chemical compound extracted from Guava Leaf, preparation method and its usage |
| CN111973612A (en) * | 2020-09-15 | 2020-11-24 | 河南大学 | Application of quercetin-3-O-beta-D-galactoside in preparation of medicament for promoting glucose absorption |
| CN111973612B (en) * | 2020-09-15 | 2023-01-24 | 河南大学 | Application of quercetin-3-O-beta-D-galactoside in preparation of medicament for promoting glucose absorption |
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